The Kit is specially designed for simultaneous purification of genomic DNA and total RNA from formalin-fixed, paraffin-embedded (FFPE) tissue sections. Purified DNA/RNA is suitable for use in applications such as real-time PCR and pyrosequencing.
Specifications
| Features | Specifications |
| Main Functions | Co-isolation DNA and RNA from FFPE tissue |
| Applications | RT-PCR, cDNA synthesis, PCR and second-generation sequencing, etc. |
| Purification method | Polydisperse silicon based magnetic beads (DNA)Monodisperse carbonyl magnetic beads (RNA) |
| Purification technology | Magnetic beads technology |
| Process method | Manual or automatic |
| Adaptive instrument | Nucleic acid extractor and pipetting workstation |
| Sample type | FFPE slice, FFPE puncture sample, embedded tissue |
| Sample amount | No more than six 10 µm sections of 150 mm2 surface area or three 20µm sections of 150 mm2 surface area |
| Yield | DNA: 1 – 10 μg, RNA: 1 – 25 μg |
The sample is lysed and digested under the action of lysate and protease. DNA/RNA is released into the lysate. After adding magnetic particles and binding solution, DNA will be adsorbed on the surface of magnetic particles, the supernatant contain RNA was collected and bind to MagBind Particles. The adsorbed particles were washed with washing solution to remove proteins and impurities, washed with ethanol to remove salts, and finally DNA/RNA was eluted by Elution Buffer.
Advantages
Kit Contents
| Contents | R632701 | R632702 | R632703 |
| Purification Times | 48 Preps | 96 Preps | 5 x 96 Preps |
| MagBind Particles | 1.1 ml | 2.5 ml | 11 ml |
| MagPure Particles N | 1.1 ml | 2.5 ml | 11 ml |
| Proteinase K | 24 mg | 48 mg | 220 mg |
| Protease Dissolve Buffer | 3 ml | 10 ml | 15 ml |
| Buffer DPS | 50 ml | 100 ml | 2 x 250 ml |
| Buffer ATL | 20 ml | 30 ml | 120 ml |
| Buffer BST1 | 20 ml | 40 ml | 200 ml |
| Buffer BXW1* | 44 ml | 110 ml | 3 x 110 ml |
| RNase Free Water | 15 ml | 30 ml | 120 ml |
Storage and Stability
Proteinase K, MagPure Particles N and MagBind Particles should be stored at 2–8°C upon arrival. However, short-term storage (up to 12 weeks) at room temperature (15–25°C) does not affect their performance. The remaining kit components can be stored at room temperature (15–25°C) and are stablefor at least 18 months under these conditions.
Experiment Data
The Kit is specially designed for simultaneous purification of genomic DNA and total RNA from formalin-fixed, paraffin-embedded (FFPE) tissue sections. Purified DNA/RNA is suitable for use in applications such as real-time PCR and pyrosequencing.
Norgen’s 2X PCR Master Mix is a ready-to-use solution that contains components required for PCR amplification including Taq DNA polymerase, dNTPs, reaction buffer, MgCl2, KCl and a PCR enhancer/stabilizer. The user needs only to add template, the primer set and water to the master mix in order to set up the PCR reaction. This convenient 2X PCR Master Mix reduces the time required to set up PCR reactions and reduces the possibility of contamination, particularly when preparing large numbers of reactions. The optimized master mix allows for robust amplification of DNA templates with high yields of PCR products.
Taq DNA Polymerase is a highly thermostable DNA polymerase that possesses a 5´→ 3´ polymerase activity and a very low 5´→ 3´ exonuclease activity. The source of Taq included with Norgen’s 2X PCR Master Mix is an E. coli strain with a cloned Taq DNA Polymerase gene from Thermus aquaticus YT-1.
Figure 1 / 2
Click for expanded view
Reagents Supplied – Cat # 28007
Storage Conditions and Product Stability
Norgen’s 2X Master Mix should be stored at -20ºC. For everyday use an aliquot can be stored at 4ºC for up to three months. The Master Mix is stable for multiple freeze-thaw cycles (see Figure 2). When stored at the proper temperature this reagent is stable for at least 1 year.
